DNA double-strand break repair of blood lymphocytes and normal tissues analysed in a preclinical mouse model: implications for radiosensitivity testing.
OBJECTIVE
Radiotherapy is an effective cancer treatment, but a few patients suffer severe radiation toxicities in neighboring normal tissues. There is increasing evidence that the variable susceptibility to radiation toxicities is caused by the individual genetic predisposition, by subtle mutations, or polymorphisms in genes involved in cellular responses to ionizing radiation. Double-strand breaks (DSB) are the most deleterious form of radiation-induced DNA damage, and DSB repair deficiencies lead to pronounced radiosensitivity. Using a preclinical mouse model, the highly sensitive gammaH2AX-foci approach was tested to verify even subtle, genetically determined DSB repair deficiencies known to be associated with increased normal tissue radiosensitivity.


METHODS
By enumerating gammaH2AX-foci in blood lymphocytes and normal tissues (brain, lung, heart, and intestine), the induction and repair of DSBs after irradiation with therapeutic doses (0.1-2 Gy) was investigated in repair-proficient and repair-deficient mouse strains in vivo and blood samples irradiated ex vivo.


RESULTS
gammaH2AX-foci analysis allowed to verify the different DSB repair deficiencies; even slight impairments caused by single polymorphisms were detected similarly in both blood lymphocytes and solid tissues, indicating that DSB repair measured in lymphocytes is valid for different and complex organs. Moreover, gammaH2AX-foci analysis of blood samples irradiated ex vivo was found to reflect repair kinetics measured in vivo and, thus, give reliable information about the individual DSB repair capacity.


CONCLUSIONS
gammaH2AX analysis of blood and tissue samples allows to detect even minor genetically defined DSB repair deficiencies, affecting normal tissue radiosensitivity. Future studies will have to evaluate the clinical potential to identify patients more susceptible to radiation toxicities before radiotherapy.
Rübe
Claudia E
CE
Department of Radiation Oncology, Saarland University, Homburg/Saar, Germany.
Grudzenski
Saskia
S
Kühne
Martin
M
Dong
Xiaorong
X
Rief
Nicole
N
Löbrich
Markus
M
Rübe
Christian
C
D000818:Animals; D001921:Brain / Q000378:metabolism / Q000528:radiation effects; D002868:Chromosomal Proteins, Non-Histone; D053903:DNA Breaks, Double-Stranded / Q000528:radiation effects*; D004260:DNA Repair / Q000502:physiology / Q000528:radiation effects*; D004268:DNA-Binding Proteins; D005455:Fluorescent Antibody Technique; D006321:Heart / Q000502:physiology / Q000528:radiation effects; D006657:Histones / Q000235:genetics / Q000378:metabolism; D007124:Immunoenzyme Techniques; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000528:radiation effects; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008168:Lung / Q000378:metabolism / Q000528:radiation effects; D008214:Lymphocytes / Q000378:metabolism / Q000528:radiation effects*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D023421:Models, Animal; D011836:Radiation Tolerance; D000071857:Tumor Suppressor p53-Binding Protein 1
D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D000071857:Tumor Suppressor p53-Binding Protein 1; C498302:Trp53bp1 protein, mouse; C475961:gamma-H2AX protein, mouse